Cloning And Expression Of Chicken Endogenous Subgroup J Avian Leukosis Gp85-like Gene

Avian leukosis virus subgroup J (ALV-J) was first isolated in 1989 and identified as a new subgroup of avian leukosis virus. The env gene of ALV-J is very unique, has very low homology to other ALV subgroups, however, exhibits 97% identity to EAV-HP sequence, which is existed in the normal chicken's genome. ALV-J most causes myeloid leukosis, while other exogenous ALVs mainly induce lymphoid leukosis. The previous research data showed that env gene is closely related to the ALV oncogenicity. Env gene code virus envelope glycoprotein, including Surface glycoprotein Unit(gp85 gene code) and transmembrane protein(gp37 gene code).Primers corresponding to the nucleotide sequences flanking the J subgroup avian leu -cosis virus( ALV-J) gp85 gene were used to amplify the intact endogenous gp85– like ge -ne from uni- nfected SPF embryo,Meat-chick embryo,Liang fenghua-chick embryo and Native-chick. The four different variety endogenous ALV-Jgp85-like elements demonstr- ated 95.2%～98.6% identical to DF1 cell Line reported, 94.9%～97.6 % identical to strain HPRS-103 ALV-J,88.5%～90.7% identical to strain IMC10200 ALV-J. For research in depth endogenous ALV-Jgp85-like express what participant action during infecting ALV-J, it pro- vides a favorable foundation.The system of Bac-to-Bac was triumphantly constructed, and expressed ALV-Jgp85-like protein. ALV-Jgp85-like protein expressed was identified by mono -clonal antibody JE9 to ALV-Jgp85 protein, by IFA and Western blot detecting. The molecular weight of ALV-Jgp85-like protein was 35KD ,as the molecular weight of ALV-Jgp85 protein(36KD) not glycosylated. The research reveals the active state of ALV-Jgp85-like in normal condition, and the immunology commonness and discrepancy with exogenous virus gp85 protein expressed.The recombined baculovirus constructed may be multipled largely, and can obtain large numbers of envelope protein with biology activity, which can be the detection antigen of ELISA instead of ALV-Jgp85 protein in the future, designed to detect the infection of ALV-J virus. The research is very significant to realize the evolution of retrovirus and the mechanism of its infection.